Abstract

BACKGROUND:

Neuroblastoma (nbl) is one of the most common solid cancers in children. Prognosis in advanced nbl is still poor despite aggressive multimodality therapy. Furthermore, survivors experience severe long-term multi-organ sequelae. Hence, the identification of new therapeutic strategies is of utmost importance. Cannabinoids and their derivatives have been used for years in folk medicine and later in the field of palliative care. Recently, they were found to show pharmacologic activity in cancer, including cytostatic, apoptotic, and antiangiogenic effects.

METHODS:

We investigated, in vitro and in vivo, the anti-nbl effect of the most active compounds in Cannabis, Δ(9)-tetrahydrocannabinol (thc) and cannabidiol (cbd). We set out to experimentally determine the effects of those compounds on viability, invasiveness, cell cycle distribution, and programmed cell death in human nbl SK-N-SH cells.

RESULTS:

Both compounds have antitumourigenic activity in vitro and impeded the growth of tumour xenografts in vivo. Of the two cannabinoids tested, cbd was the more active. Treatment with cbd reduced the viability and invasiveness of treated tumour cells in vitro and induced apoptosis (as demonstrated by morphology changes, sub-G1 cell accumulation, and annexin V assay). Moreover, cbd elicited an increase in activated caspase 3 in treated cells and tumour xenografts.

CONCLUSIONS:

Our results demonstrate the antitumourigenic action of cbd on nbl cells. Because cbd is a nonpsychoactive cannabinoid that appears to be devoid of side effects, our results support its exploitation as an effective anticancer drug in the management of nbl.

Δ9-Tetrahydrocannabinol (THC) and cannabidiol (CBD) reduce viability of neuroblastoma (NBL) cell lines in vitro, with CBD having a better effect. (A) Cell lines SK-N-SH (open squares), NUB-6 (open circles), IMR-32 (open triangles), and LAN-1 (crosses) were incubated with increasing concentrations (0–50 μg/mL) of THC or CBD for 24 hours and 48 hours. Viability was measured by MTT assay. (B) Mean ± standard deviation of SK-N-SH cell viability after incubation with 10 μg/mL THC or CBD for 24 and 48 hours. *** Denotes a significant change relative to control (p = 0.0004). Data are expressed as a percentage of the vehicle control and are the mean of pooled results from experiments performed in triplicate.

Anti-invasiveness effect of cannabidiol (CBD) on SK-N-SH cells. The invasion assays were performed using cell cultures (2×105 cells/well) treated with CBD (15 μg/mL, 20 μg/mL) for 24 hours; results were compared with those for untreated cells (2×105 cells/well). For each well (treated or untreated cells), 10 fields were examined by light microscopy.